Molecular and Biochemical Analysis of Chalcone Synthase from Freesia hybrid in flavonoid biosynthetic pathway.
Molecular and Biochemical Analysis of Chalcone Synthase from Freesia hybrid in flavonoid biosynthetic pathway.
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小苍兰杂种黄酮生物合成途径中查尔酮合酶的分子和生化分析
DOI：
10.1371/journal.pone.0119054
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Wang L
中科院分区：
文献类型：
--
作者：
Sun W;Meng X;Liang L;Jiang W;Huang Y;He J;Hu H;Almqvist J;Gao X;Wang L
查耳酮合酶（CHS）催化类黄酮生物合成途径中的第一个关键步骤。本研究成功地从小苍兰中克隆了编码CHS的cDNA（FhCHS1）。多重序列比对结果表明，FhCHS1的氨基酸序列中存在保守的CHS活性位点残基和CHS特征序列。同时，晶体学分析表明FhCHS1与苜蓿CHS2的蛋白质结构高度相似，生化分析结果表明FhCHS1在柚皮素生物合成中具有酶促作用。此外，利用实时荧光定量PCR技术检测了FhCHS1在花和不同组织中的转录水平，FhCHS1在花中的表达模式与花发育过程中花色素苷的积累模式存在显著相关性。为了进一步表征FhCHS1的功能，进行了其在拟南芥tt4突变体和矮牵牛中的异位表达。结果表明，在tt4突变体中过量表达FhCHS1完全恢复了种皮、子叶和下胚轴的色素表型，而表达FhCHS1的转基因矮牵牛的花色从白色变为粉红色。综上所述，这些结果表明，FhCHS1在小苍兰类黄酮的生物合成中起着至关重要的作用，并可用于修饰其他植物中的类黄酮成分。
Chalcone synthase (CHS) catalyzes the first committed step in the flavonoid biosynthetic pathway. In this study, the cDNA (FhCHS1) encoding CHS from Freesia hybrida was successfully isolated and analyzed. Multiple sequence alignments showed that both the conserved CHS active site residues and CHS signature sequence were found in the deduced amino acid sequence of FhCHS1. Meanwhile, crystallographic analysis revealed that protein structure of FhCHS1 is highly similar to that of alfalfa CHS2, and the biochemical analysis results indicated that it has an enzymatic role in naringenin biosynthesis. Moreover, quantitative real-time PCR was performed to detect the transcript levels of FhCHS1 in flowers and different tissues, and patterns of FhCHS1 expression in flowers showed significant correlation to the accumulation patterns of anthocyanin during flower development. To further characterize the functionality of FhCHS1, its ectopic expression in Arabidopsis thaliana tt4 mutants and Petunia hybrida was performed. The results showed that overexpression of FhCHS1 in tt4 mutants fully restored the pigmentation phenotype of the seed coats, cotyledons and hypocotyls, while transgenic petunia expressing FhCHS1 showed flower color alteration from white to pink. In summary, these results suggest that FhCHS1 plays an essential role in the biosynthesis of flavonoid in Freesia hybrida and may be used to modify the components of flavonoids in other plants.
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DOI：
10.1073/pnas.93.17.9033
发表时间：
1996-08-20
影响因子：
11.1
作者：
Helariutta, Y;Kotilainen, M;Albert, VA
通讯作者：
Albert, VA
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
7.2
作者：
Clough, SJ;Bent, AF
通讯作者：
Bent, AF
影响因子：
5.2
作者：
Abe, I;Takahashi, Y;Noguchi, H
通讯作者：
Noguchi, H
DOI：
10.1038/11553
发表时间：
1999-08-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Ferrer, JL;Jez, JM;Noel, JP
通讯作者：
Noel, JP